J. Parasitol., 89(5), 2003, pp. 1039±1043 ᭧ American Society of Parasitologists 2003

TWO OF ACUARIA BREMSER, 1811 (NEMATODA: ACUARIOIDEA: ACUARIIDAE) IN PASSERINE FROM THE AREA DE CONSERVACION GUANACASTE, COSTA RICA

Luping Zhang, Daniel R. Brooks*, and Douglas Causey† Department of Zoology, University of Toronto, Ontario, Canada M5S 3G5. e-mail: [email protected]

ABSTRACT: Two species of Acuaria were collected from passerine birds from the Area de Conservacion Guanacaste, Costa Rica. Acuaria mayori Lent, Freitas and Proenca, 1945, was collected from Myiarchus nuttingi. Specimens from Costa Rica differ from the original description by having a spicule ratio of 1:1.5±1.7 versus 1:1.43±1.47, as well as shorter spicules and female tails. Acuaria wangi n. sp. in Hylophylax naevioides and Gymnopithys leucaspis resembles A. alii, A. crami, A. cyanocitta, A. minuta, A. pattoni, and A. cissae by having cordons extending posteriorly to the anterior portion of the glandular esophagus. The new species differs from A. alii by having 4 pairs of preanal and 6 pairs of postanal papillae rather than 2 pairs of preanal and 7 pairs of postanal papillae, a shorter left spicule, a spicule ratio of 1:1.6±1.8 versus 1:1.1 and in having spicules with blunt rather than pointed distal ends. Acuaria crami and A. minuta differ from A. wangi by having 7 pairs of postanal papillae and spicule ratios of 1:1.6±1.8 versus 1:1.3 in A. crami and 1:1.1 in A. minuta; in addition, A. minuta has spatulate-shaped spicules and a tricupsid-shaped distal end of the right spicule. The new species can be distinguished from A. pattoni by having a longer left spicule and a spicule ratio of 1:1.6±1.8 versus 1:1 and from A. cissae by having a shorter left spicule and a spicule ratio of 1: 1.6±1.8 versus 1:2.5±2.7. Acuaria wangi is similar to A. cyanocitta, which has similarly shaped spicules, including a very pointed distal end of the left spicule, but differs in body length, in having shorter spicules, in the arrangement of postanal papillae, and in having smaller eggs.

Acuaria Bremser, 1811, is the most species-rich of DESCRIPTION acuariid , with more than 70 nominal species (Sher- Acuaria mayori Lent, Freitas and Proenca, 1945 win and Schmidt, 1988). All but 1 known species inhabit pas- (Figs. 1–6) serine birds, the exception being A. upupa Rasheed, 1960, from the coraciiform , Upupa epops, from India (Mawson, 1972). Diagnosis: Body small with transverse striations. Two pseu- Bremser (1811) proposed Acuaria without designating a type dolabia lateral to mouth, each bearing a pair of large cephalic species. Of the 14 species he included in Acuaria, Spiroptera papillae and 1 inconspicuous amphid. Cordons straight, non- anthuris Rudolphi, 1819, is regarded as the type species for anastomosing, and nonrecurrent, originating at dorsal and ven- Anthuris Rudolphi, 1819, by absolute tautonomy and by Ru- tral sides of oral opening, continuing posteriorly to middle of dolphi's original intentions, but Rudolphi admitted that Anthuris muscular esophagus, 3.2±4.8% (4.0%) total body length (TBL) was Acuaria Bremser, 1811, renamed; hence, the type species in male and 1.9±2.7% (2.4%) TBL in female. Buccal capsule A. anthuris is the type species of Acuaria (Cram, 1927). Skrja- long and slender, transversely striated. Esophagus clearly divid- bin et al. (1965) regarded Cheilospirura Diesing, 1861, as a ed into short anterior muscular part and long posterior glandular synonym of Acuaria, but Chabaud (1975) considered Cheilos- part. Muscular esophagus 4.0±5.6% (5.2%) TBL in male and pirura a valid genus. 2.2±3.4% (2.7%) TBL in female; glandular esophagus 9.8± During June 2001 and January 2002, as part of an ongoing 12.7% (11.4%) TBL in male and 4.8±5.8% (5.2%) TBL in fe- biodiversity inventory of the eukaryotic parasites of vertebrates male. Nerve ring located at the level of the anterior end of the inhabiting the Area de Conservacion Guanacaste (ACG) in muscular esophagus. Cervical papillae located at the same level as nerve ring. Excretory pore immediately posterior to nerve northwestern Costa Rica (http://brooksweb.zoo.utoronto.ca/ ring. index.html), 2 species of Acuaria were collected from passerine Male (n ϭ 7): Body length 5.427±7.452 mm (6.808 mm). birds. We describe both in this article. Maximum width 111±148 (131). Cordons 239±323 (275) long. Buccal capsule 114±156 (134) long. Muscular esophagus 266± MATERIALS AND METHODS 429 (355) long and 30±38 (34) wide; glandular esophagus 646± Nutting's ¯ycatchers, Myiarchus nuttingi Ridgway, 1883 (n ϭ 5), 893 (778) long and 49±61 (56) wide. Nerve ring 171±209 (188) spotted antbirds, Hylophylax naevioides (Lafresnaye, 1847) (n ϭ 23), from anterior end, excretory pore 255±289 (268) from anterior and bicolored antbirds, Gymnopithys leucaspis (Sclater, 1885) (n ϭ 7) end, and cervical papillae 167±213 (188) from anterior end. were collected in June 2001 and January 2002 and examined for par- Posterior end of body curved. Caudal alae well developed, 311± asites. Nematodes collected from beneath the lining of the gizzard were ®xed in glacial acetic acid and preserved in 70% ethanol. They were 396 (348) long, and 38±60 (48) wide. Tail bluntly rounded, later cleared in lactophenol for further examination. Drawings were 162±178 (169) long. Eleven pairs of caudal papillae, 4 pairs made with the aid of a drawing tube. Measurements (range, followed preanal and 7 pairs postanal. Spicules subequal and similar, by mean in parentheses) are given in micrometers unless otherwise with rounded distal end. Left spicule 163±186 (179) long, 12.9± stated. 16.1 (15.9) wide at base of proximal end. Right spicule 102± 123 (111) long, 12.9±19.3 (16.1) wide at base of proximal end. Ratio of right spicule±left spicule 1:1.5±1.7 (1:1.6). Received 9 December 2002; revised 7 April 2003; accepted 7 April Female (n ϭ 7): Body length 13.84±22.03 mm (18.74 mm). 2003. * To whom correspondence should be addressed. Maximum width 163±192 (173). Cordons 368±518 (444) long. ² Museum of Comparative Zoology, Harvard University, Cambridge, Buccal capsule 143±182 (159) long. Muscular esophagus 441± Massachusetts. 597 (509) long and 32±57 (45) wide; glandular esophagus 799±

1039 1040 THE JOURNAL OF PARASITOLOGY, VOL. 89, NO. 5, OCTOBER 2003

FIGURES 1±6. Acuaria mayori Lent, Freitas and Proenca, 1945. 1. Anterior region of female, lateral view. 2±3. Posterior end of male, ventral view. 4. Spicules. 5. Posterior end of female, lateral view. 6. Vulva region of female, lateral view. Bars ϭ 50 ␮m. ZHANG ET AL.—TWO SPECIES OF ACUARIA IN COSTA RICAN BIRDS 1041

1,254 (975) long and 48±87 (74) wide. Nerve ring 200±247 (273) from anterior end, and cervical papillae 148±171 (159) (227) from anterior end, excretory pore 292±353 (329) from from anterior end. Caudal alae well developed, 247±285 (271) anterior end, and cervical papillae 205±258 (227) from anterior long, and 32±37 (34) wide. Tail bluntly rounded, 133±152 (143) end. Didelphic. Vulva near the middle of body, 6.74±10.37 mm long. Ten pairs of caudal papillae, 4 pairs of preanal and 6 pairs (9.31 mm) from anterior end, 46±58.4% (49.6%) TBL from of postanal papillae (1 male had only 4 pairs of postanal pa- anterior end. Tail short and rounded, 129±179 (148) long. Eggs pillae) (Fig. 9). Postanal papillae divided into 2 groups, each ellipsoid, thick shelled, embryonated, 31.3±38.6 (34.0) long by group includes 3 pairs of papillae (Fig. 8). Spicules subequal 18.4±23.9 (20.6) wide. but dissimilar. Left spicule long and slender, 190±209 (199) long, 9.7±12.9 (10.9) wide at base of proximal end, with a Taxonomic summary pointed distal end. Right spicule short and thick, 103±129 (116) long, 16.1±22.5 (18.9) wide at base of proximal end, with a Host: Myiarchus nuttingi (Aves: Passeriformes: Tyrannidae). rounded distal end. Ratio of right spicule:left spicule 1:1.6±1.8 Localities: Cafetal, Sector Santa Rosa, ACG, Guanacaste (1:1.7). Province, Costa Rica, 10Њ51Ј19ЉN, 85Њ36Ј39ЉW. Quebrada Costa Female (n ϭ 9): Body length 6.80±10.04 mm (8.59 mm). Rica, Sector Santa Rosa, ACG, Guanacaste Province, Costa Maximum width 205±311 (251). Cordons 1.26±1.85 mm (1.55 Rica; 10Њ49Ј39ЉN, 85Њ38Ј12ЉW, new locality. mm) long. Buccal capsule 144±190 (162) long. Muscular Site of infection: Under the lining of the gizzard. esophagus 623±844 (711) long and 49±65 (53) wide; glandular Prevalence: Forty percent (2 of 5 birds). esophagus 1.48±1.88 mm (1.69 mm) long and 72±103 (83) Intensity: Two males and 9 females in 1 host, and 5 males wide. Nerve ring 197±266 (224) from anterior end, excretory and 1 female in the second host. pore 293±380 (324) from anterior end, and cervical papillae Voucher specimens: USNPC 93584, 93585. 171±228 (201) from anterior end. Didelphic. Vulva just poste- rior to middle of body, 3.56±5.64 mm (4.63 mm) from anterior Remarks end, 51±57% (54%) TBL from anterior end. Tail short and Acuaria mayori Lent, Freitas and Proenca, 1945, was ®rst rounded, 141±179 (157) long. Eggs ellipsoid, thick shelled, em- described in Cyanocorax chrysops (Vieillot, 1818) from Para- bryonated, 31±37 (36) long by 18±22 (20) wide. guay and has since been reported in Sporophila caerulescens caerulescens (Vieillot, 1823) and Cyanocorax cyanomelas Taxonomic summary (Wied, 1821) from Brazil (Pinto et al., 1993, 1997). The present specimens are similar to A. mayori in cordon length, number Type host: Hylophylax naevioides (Aves: Passeriformes: For- and arrangement of male caudal papillae, and spicule shape but micariidae). differs from the original description and previous reports by Other host: Gymnopithys leucaspis (Aves: Passeriformes: having a spicule ratio of 1:1.5±1.7 versus 1:1.43±1.47, as well Formicariidae). as shorter spicules (left spicule 163±186 vs. 190±230; right Type locality: EstacioÂn San Gerardo, San Gerardo, ACG, spicule 102±123 vs. 130±160) and female tails (129±179 vs. Guanacaste Province, Costa Rica; 10Њ52Ј50ЉN, 85Њ23Ј21ЉW. 170±230). This is the ®rst report of A. mayori from Costa Rica, Site of infection: Under the lining of the gizzard. and M. nuttingi is a new host record. Prevalence: In H. naevioides, 17.4% (4 of 23 birds). In G. leucaspis, 14.3% (1 of 7 birds). Intensity: Range in H. naevioides,1±4;inG. leucaspis,2. Acuaria wangi n. sp. (Figs. 7–11) Type specimens: Holotype, USNPC 93544; allotype, USNPC 93545; paratypes, USNPC 93546, 93547, 93548, 93549, 93550. Diagnosis: Body small with transverse striations. Two pseu- Etymology: The new species is named after Professor Puqing dolabia lateral to mouth, each bearing a pair of large cephalic Wang, Laboratory of Parasitology, Fujian Normal University, papillae and 1 inconspicuous amphid. Cordons straight, non- People's Republic of China, for his contribution to the fauna anastomosing, and nonrecurrent, originating at dorsal and ven- and of nematodes. tral sides of oral opening, continuing posteriorly to anterior part of glandular esophagus, 15.5±19.8% TBL. Buccal capsule long Remarks and slender, transversely striated. Esophagus clearly divided into short anterior muscular part and long posterior glandular By having cordons extending posteriorly to the anterior por- part. Muscular esophagus 10.5±11.5% (11%) TBL in male and tion of the glandular esophagus, A. wangi resembles A. alii 6.7±10.3% (8.3%) TBL in female; glandular esophagus 26.5± Rasheed, 1960, in Acridotheres ginginianus (Latham, 1790) 29.3% (28.1%) TBL in male and 15.5±27.7% (19.7%) TBL in from India, A. crami Rasheed, 1960, in Saxicola caprata ca- female. Nerve ring located at level of anterior end of muscular prata (Linnaeus, 1766) from India (probably not the correct esophagus. Cervical papillae immediately anterior to nerve host identi®cation as S. c. caprata occurs only in Luzon, Phil- ring. Excretory pore posterior to nerve ring. ippines), A. cyanocitta (Boyd, 1956) in Cyanocitta cristata Male (n ϭ 4): Body length 4.212±4.884 mm (4.645 mm). (Linnaeus, 1758) from the United States, A. minuta Williams, Maximum width 133±185 (170). Cordons 786±951 (868) long. 1929, in Quiscalus quiscala aeneus (this host record is also Buccal capsule 137±156 (146) long. Muscular esophagus 486± doubtful because Q. q. aenus (Linnaeus, 1758) is not a valid 524 (511) long and 30±53 (41) wide; glandular esophagus name and authority combination; Q. q. quiscala (L., 1758) is 1.159±1.425 mm (1.306 mm) long and 76±99 (86) wide. Nerve the type by tautonomy, and Q. aeneus Ridgway, 1869, has been ring 182±205 (198) from anterior end, excretory pore 263±285 synonymized with Q. q. versicolor (Vieillot, 1819) from the 1042 THE JOURNAL OF PARASITOLOGY, VOL. 89, NO. 5, OCTOBER 2003

FIGURES 7±11. Acuaria wangi n. sp. 7. Anterior region of female, lateral view. 8±9. Posterior end of male, ventral view. 10. Posterior end of female, lateral view. 11. Vulva region of female, lateral view. Bars ϭ 50 ␮m.

United States, A. pattoni Williams, 1929, in Sturnella neglecta extending behind the glandular esophagus or to the posterior (Audubon, 1844) from the United States, and A. cissae Wang, part of body. 1976, in Cissa e. erythrorhyncha (Boddaert, 1783) from China. The new species, however, differs from A. alii in having 4 Other members of the genus either have very short cordons, pairs of preanal and 6 pairs of postanal rather than 2 pairs of extending to the part of muscular esophagus, or longer cordons, preanal and 7 pairs of postanal papillae, a longer left spicule ZHANG ET AL.—TWO SPECIES OF ACUARIA IN COSTA RICAN BIRDS 1043

(190±209 vs. 110), a different spicule ratio (1:1.6±1.8 vs. 1: were made by D.R.B., D.C., Elda Araya, Sara Brant, Marie Causey, 1.1), and a right spicule with a blunt rather than pointed distal Ben Hanelt, Calixto Moraga, and Petrona Rios. This study was funded by a research grant from the Natural Sciences and Engineering Research end. The new species can be distinguished from A. crami and Council of Canada to D.R.B. and by a grant from the MCZ Putnam A. minuta by its possession of 6 pairs of postanal papillae and Expedition Fund to D.C. a different spicule ratio (1:1.6±1.8 vs. 1:1.3 in A. crami and 1: 1.1 in A. minuta); in addition, A. minuta has spatulate-shaped LITERATURE CITED spicules and a tricuspid-shaped distal end of the right spicule. BREMSER, J. G. 1811. Nachricht von einer betrachtlichen Sammlung The remaining 3 species resemble A. wangi by possessing 4 thierischer Eingeweidewurmer. etc. k. k. Naturalienkabinets. Direk- preanal and 6 postanal papillae. Of these, the new species can tion in Wien. S. 51. be distinguished from A. pattoni by having a longer left spicule CHABAUD, A. G. 1975. Keys to the genera of the order . Part (190±209 vs. 156) and a spicule ratio of 1:1.6±1.8 versus 1:1 2. Spiruroidea, Habronematoidea and Acuarioidea. In CIH keys to the parasites of vertebrates. No. 3, R. C. Anderson, A. and from A. cissae by having a shorter left spicule (190±209 G. Chabaud, and S. Willmott (eds.). Commonwealth Agricultural vs. 268±280) and a spicule ratio of 1:1.6±1.8 versus 1:2.5±2.7. Bureaux, Farnham Royal, U.K., p. 29±58. Acuaria wangi is related to A. cyanocitta, which has similarly ÐÐÐ, AND A. PETTER. 1961. Nematodes du genre Acuaria de la faune shaped spicules, including a very pointed distal end of the left de France. Annales de Parasitologie humaine et Comparee 36: 409±424. spicule and a similar spicule ratio (1:1.6±1.8 vs. 1:1.4±1.6), but CRAM, E. 1927. Bird parasites of the nematode suborders Strongylata, the new species differs in body length (male 4.2±4.9 mm long Ascaridata and Spirurata. United States National Museum Bulletin and female 6.8±10.0 mm long in A. wangi vs. male 7.5±10.0 140: 1±465. mm long and female 12.8±18.4 mm long in A. cyanocitta), in GUPTA,S.P.,AND M. JEHAN. 1972. On some species of the genus Ac- having shorter spicules (left spicule 190±209 vs. 315±370; right uaria Bremser, 1811 from avian hosts from Lucknow. Japanese Journal of Parasitology 21: 365±373. spicule 103±129 vs. 220±230), in the arrangement of postanal ÐÐÐ, AND P. K UMAR. 1977. On some species of the genus Acuaria papillae (2 groups vs. 3 groups), and in having smaller eggs Bremser, 1811 from avian hosts from Uttar Prasesh. Indian Journal (31±37 long by 18±22 wide vs. 42 long by 24 wide). of Helminthology 29: 120±136. HSU, W. N. 1963. Studies on some parasitic nematodes of order Spi- DISCUSSION rurida Chitwood, 1933. Acta Zoologica Sinica 15: 544±552. [In Chinese.] Acuaria mayori was collected only from dry forest sites (Caf- ILYAS, R. 1980. On a new species of Acuaria Bremser, 1811 from Gul- lus domesticus. Revista di Parasitologia 41: 73±76. etal, Quebrada Costa Rica), whereas A. wangi occurred in a wet MAWSON, P. M. 1972. The genus Acuaria Bremser (Nematoda: Spiru- forest site (EstacioÂn San Gerardo). These are sites with abun- rida) in Australia. Transactions of the Royal Society of South Aus- dant running water and associated riparian vegetation. Acuaria tralia 96: 139±147. mayori was collected at the beginning of the wet season (5 and PINTO, R. M., J. J. VICENTE, AND D. NORONHA. 1997. Nematode parasites 7 June 2001), whereas A. wangi was collected at the beginning of Brazilian corvid birds (Passeriformes): A general survey with a description of Viktorocara braziliensis n. sp. (Acuariidae, Schis- of the wet season (13±14 June 2001 in H. naevioides and G. torophinae). Memorias do Instituto Oswaldo Cruz 92: 209±214. leucaspis) and at the beginning of the dry season (13 January ÐÐÐ, ÐÐÐ, ÐÐÐ, AND S. P. DE FABIO. 1993. New records of the 2002 in H. naevioides). nematodes Ascaridia columbae (Gmelin) Travassos, Acuaria may- ori Lent, Freitas and Proenca and Aproctella stoddardi Cram in ACKNOWLEDGMENTS Brazilian birds, with redescription of the species. Revista Brazileira de Zoologia 8: 1±6. We are grateful to the scienti®c and technical staff of the ACG for SHERWIN,F.J.,AND G. D. SCHMIDT. 1988. Helminths of the Mountains supporting this study, in particular, Elda Araya, Roger Blanco, Carolina of Colorado, including Acuaria coloradensis n. sp. Journal of Par- Cano, Maria Marta ChavarrõÂa, Felipe ChavarrõÂa, Roberto Espinoza, asitology 74: 336±338. Dunia Garcia, Guillermo Jimenez, Elba Lopez, Sigifredo Marin, Ale- SKRJABIN, K. I., A. A. SOBOLEV, AND V. M. IVASHKIN. 1965. Principles jandro Masis, Calixto Moraga, Fredy Quesada, and Petrona Rios. of Nematology 14. Spirurata of and man and the diseases Thanks are also due to Dan Janzen and Winnie Hallwachs, scienti®c caused by them. Part 3. Acuarioidea Izdatel'stvo Nauka, Moscow, advisers to the ACG, for their support. Host specimens were collected Russia. p. 31±132. [In Russian.] by D.C., Jeremiah Trimble (Museum of Comparative Zoology (MCZ)), WANG, P. 1976. Notes on some new nematodes of suborder Spirurata and Calixto Moraga (ACG) under the authority of CITES (Commission from Fujian, China. Acta Zoologica Sinica 22: 393±402. [In Chi- on International Trade in Endangered Species) Permit US9258251, nese.] CITES Permit CR9123440, Costa Rica Ministero del Ambiente y Ener- ZHANG, L. P. 1990. Three new species of Acuarioidea from birds from gia Licencia 203640283 and Resoluciones 215-2001-OFAU and 411- Fujian, China. Sichuan Journal of Zoology 9: 1±5. [In Chinese.] 2001-OFAU, Harvard University IACUC Protocol 21-09, and U.S. De- ÐÐÐ, AND Z. Y. WANG. 1993. A new species of Acuarioides from partment of Agriculture and Plant Health Inspection Service birds (Spirurida: Acuariidae). Sichuan Journal of Zoology 12: 1± Permit 47956 (Form VS16-6A). Host necropsy and parasite collections 3. [In Chinese.]